Analogues of GLP-1

ABSTRACT

Disclosed are peptide analogues of glucagon-like peptide-1, the pharmaceutically-acceptable salts thereof, methods of using such analogues to treat mammals and pharmaceutical compositions useful therefor comprising said analogues.

BACKGROUND OF THE INVENTION

This application is a United States national stage filing under 35U.S.C. §371 of international (PCT) application No. PCT/US2004/042045,filed Dec. 15, 2004 and designating the US, which application claimspriority to U.S. provisional application 60/529,822 filed Dec. 16, 2003.

The present invention is directed to peptide analogues of glucagon-likepeptide-1, the pharmaceutically-acceptable salts thereof, to methods ofusing such analogues to treat mammals and to pharmaceutical compositionsuseful therefor comprising said analogues.

Glucagon-like peptide-1 (7-36) amide (GLP-1) is synthesized in theintestinal L-cells by tissue-specific post-translational processing ofthe glucagon precursor preproglucagon (Varndell, J. M., et al., J.Histochem Cytochem, 1985:33:1080-6) and is released into the circulationin response to a meal. The plasma concentration of GLP-1 rises from afasting level of approximately 15 pmol/L to a peak postprandial level of40 pmol/L. It has been demonstrated that, for a given rise in plasmaglucose concentration, the increase in plasma insulin is approximatelythreefold greater when glucose is administered orally compared withintravenously (Kreymann, B., et al., Lancet 1987:2, 1300-4). Thisalimentary enhancement of insulin release, known as the incretin effect,is primarily humoral and GLP-1 is now thought to be the most potentphysiological incretin in humans. In addition to the insulinotropiceffect, GLP-1 suppresses glucagon secretion, delays gastric emptying(Wettergren A., et al., Dig Dis Sci 1993:38:665-73) and may enhanceperipheral glucose disposal (D'Alessio, D. A. et al., J. Clin Invest1994:93:2293-6).

In 1994, the therapeutic potential of GLP-1 was suggested following theobservation that a single subcutaneous (s/c) dose of GLP-1 couldcompletely normalize postprandial glucose levels in patients withnon-insulin-dependent diabetes mellitus (NIDDM) (Gutniak, M. K., et al.,Diabetes Care 1994:17:1039-44). This effect was thought to be mediatedboth by increased insulin release and by a reduction in glucagonsecretion. Furthermore, an intravenous infusion of GLP-1 has been shownto delay postprandial gastric emptying in patients with NIDDM (Williams,B., et al., J. Clin Endo Metab 1996:81:327-32). Unlike sulphonylureas,the insulinotropic action of GLP-1 is dependent on plasma glucoseconcentration (Holz, G. G. 4^(th), et al., Nature 1993:361:362-5). Thus,the loss of GLP-1-mediated insulin release at low plasma glucoseconcentration protects against severe hypoglycemia. This combination ofactions gives GLP-1 unique potential therapeutic advantages over otheragents currently used to treat NIDDM.

Numerous studies have shown that when given to healthy subjects, GLP-1potently influences glycemic levels as well as insulin and glucagonconcentrations (Orskov, C, Diabetologia 35:701-711, 1992; Hoist, J. J.,et al., Potential of GLP-1 in diabetes management in Glucagon III,Handbook of Experimental Pharmacology, Lefevbre P J, Ed. Berlin,Springer Verlag, 1996, p. 311-326), effects which are glucose dependent(Kreymann, B., et al., Lancet ii:1300-1304, 1987; Weir, G. C., et al.,Diabetes 38:338-342, 1989). Moreover, it is also effective in patientswith diabetes (Gutniak, M., N. Engl J Med 226:1316-1322, 1992; Nathan,D. M., et al., Diabetes Care 15:270-276, 1992), normalizing bloodglucose levels in type 2 diabetic subjects (Nauck, M. A., et al.,Diagbetologia 36:741-744, 1993), and improving glycemic control in type1 patients (Creutzfeldt, W. O., et al., Diabetes Care 19:580-586, 1996),raising the possibility of its use as a therapeutic agent.

GLP-1 is, however, metabolically unstable, having a plasma half-life(t_(1/2)) of only 1-2 min in vivo. Exogenously administered GLP-1 isalso rapidly degraded (Deacon, C. F., et al., Diabetes 44:1126-1131,1995). This metabolic instability limits the therapeutic potential ofnative GLP-1. Hence, there is a need for GLP-1 analogues that are moreactive and/or are more metabolically stable than native GLP-1.

SUMMARY OF THE INVENTION

In one aspect the invention features a compound according to formula(I),(R2R3)-A⁷-A⁸-A⁹-A¹⁰-A¹¹-A¹²-A¹³-A¹⁴-A¹⁵-A¹⁶-A¹⁷-A¹⁸-A¹⁹-A²⁰-A²¹-A²²-A²³-A²⁴-A²⁵-A²⁶-A²⁷-A²⁸-A²⁹-A³⁰-A³¹-A³²-A³³-A³⁴-A³⁵-A³⁶-A³⁷-A³⁸-A³⁹-R¹,  (I)wherein

-   A⁷ is L-His, Ura, Paa, Pta, Amp, Tma-His, des-amino-His, or deleted;-   A⁸ is Ala, β-Ala, Gly, Ser, D-Ala, Aib, Acc, N-Me-Ala, N-Me-D-Ala or    N-Me-Gly;-   A⁹ is Glu, N-Me-Glu, N-Me-Asp or Asp;-   A¹⁰ is Gly, Acc, β-Ala or Aib;-   A¹¹ is Thr or Ser;-   A¹² is Phe, Acc, Aic, Aib, 2-Pal, 3-Pal, 4-Pal, 1Nal, 2Nal, Cha, Trp    or (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe;-   A¹³ is Thr or Ser;-   A¹⁴ is Ser or Aib;-   A¹⁵ is Asp or Glu;-   A¹⁶ is Val, Acc, Aib, Leu, Ile, Tle, Nle, Abu, Ala or Cha;-   A¹⁷ is Ser, Aib or Thr;-   A¹⁸ is Ser, Lys or Thr;-   A¹⁹ is Tyr, Cha, Phe, 2-Pal, 3-Pal, 4-Pal, 1Nal, 2Nal, Acc or    (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe;-   A²⁰ is Leu, Acc, Aib, Nle, Ile, Cha, Tle, Val, Phe or    (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe;-   A²¹ is Glu or Asp;-   A²² is Gly, Acc, β-Ala, Glu or Aib;-   A²³ is Gln, Asp, Asn or Glu;-   A²⁴ is Ala, Aib, Val, Abu, Tle or Acc;-   A²⁵ is Ala, Aib, Val, Abu, Tle, Acc, Lys, Arg, hArg, Orn,    HN—CH((CH₂)_(n)—N(R¹⁰R¹¹))—C(O) or HN—CH((CH₂)_(e)—X³)—C(O);-   A²⁶ is Lys, Arg, hArg, Orn, Lys(N^(ε)-decanoyl)),    HN—CH((CH₂)_(n)—N(R¹⁰R¹¹))—C(O) or HN—CH((CH₂)_(e)—X³)—C(O);-   A²⁷ is Glu, Asp, Leu, Aib or Lys;-   A²⁸ is Phe, 2-Pal, 3-Pal, 4-Pal, 1Nal, 2Nal, (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe,    Aic, Acc, Aib, Cha or Trp;-   A²⁹ is Ile, Acc, Aib, Leu, Nle, Cha, Tle, Val, Abu, Ala or Phe;-   A³⁰ is Ala, Aib or Acc;-   A³¹ is Trp, 2-Pal, 3-Pal, 4-Pal, 1Nal, 2Nal, Phe, Acc, Aib,    (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe or Cha;-   A³² is Leu, Acc, Aib, Nle, Ile, Cha, Tle, Phe, (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe    or Ala;-   A³³ is Val, Acc, Aib, Leu, Ile, Tle, Nle, Cha, Ala, Phe, Abu, Lys or    (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe;-   A³⁴ is Lys, Arg, hArg, Orn, HN—CH((CH₂)_(n)—N(R¹⁰R¹¹))—C(O) or    HN—CH((CH₂)_(e)—X³)—C(O);-   A³⁵ is β-Ala, D-Ala, Gaba, Ava, HN—(CH₂)_(m)—C(O), Aib, Acc, D-Arg    or a D-amino acid;-   A³⁶ is L- or D-Arg, D- or L-Lys, or Lys(N^(ε)-decanoyl) or    Lys(N^(ε)-dodecanoyI) or D- or L-hArg, D- or L-Orn or    HN—CH((CH₂)_(n)—N(R¹⁰R¹¹))—C(O), or HN—CH((CH₂)_(e)—X³)—C(O);-   A³⁷ is Gly, β-Ala, Gaba, Aib, Acc, Act, Apc, Aun, Ava, Pro, Dhp,    Dmt, Pip, L- or D-Arg, L- or D-Asp or Glu, Lys(N^(ε)-decanoyl),    Lys(N^(ε)-dodecanoyl), Lys(N^(ε)-octanoyl),    Lys(N^(ε)-tetradecanoyl), or Ser(O-decanoyl);-   A³⁸ is D- or L-His, L- or D-Ala, Asn, Gln, Ser, Thr, Acc, Ado, Aib,    Apc, Act, Arg, Ava, Gly, β-Ala, Gaba, or HN—(CH₂)_(s)—C(O);-   A³⁹ is D- or L-His, L- or D-Ala, Asn, Gln, Ser, Thr, Acc, Ado, Aib,    Apc, Act, Arg, Aun, Gly, β-Ala, Gaba, Lys(N^(ε)-octanoyl),    HN—(CH₂)_(s)—C(O), or deleted;-   R¹ is OH, NH₂; (C₁-C₃₀)alkoxy, or NH—X²—CH₂—Z⁰, wherein X² is a    (C₀-C₂), (C₄-C₉) or (C₁₁-C₁₉)hydrocarbon moiety and Z⁰ is H, OH,    CO₂H or CONH₂;

or —C(O)—NHR¹², wherein X⁴ is, independently for each occurrence,—C(O)—, —NH—C(O)— or —CH₂—, and wherein f is, independently for eachoccurrence, an integer from 1 to 29 inclusive;

-   each of R² and R³ is independently selected from the group    consisting of H, (C₁-C₃₀)alkyl, (C₂-C₃₀)alkenyl, optionally    substituted phenyl(C₁-C₃₀)alkyl, optionally substituted    naphthyl(C₁-C₃₀)alkyl, hydroxy(C₁-C₃₀)alkyl, hydroxy(C₂-C₃₀)alkenyl,    hydroxyphenyl(C₁-C₃₀)alkyl, and hydroxynaphthyl(C₁-C₃₀)alkyl;

wherein the phenyl group of said optionally substitutedphenyl(C₁-C₃₀)alkyl moiety, and said naphthyl group of said optionallysubstituted naphthyl(C₁-C₃₀)alkyl moiety each is, independently for eachoccurrence, substituted with 1 or more substituents selected,independently for each occurrence, from the group consisting of halo,OH, NH₂, NO₂ and CN;

-   or one of R² and R³ is

(C₁-C₃₀)acyl, (C₁-C₃₀)alkylsulfonyl, C(O)X⁵,

wherein Y is H, OH or NH₂; r is 0 to 4; q is 0 to 4; and X⁵ is(C₁-C₃₀)alkyl, (C₂-C₃₀)alkenyl, phenyl(C₁-C₃₀)alkyl,naphthyl(C₁-C₃₀)alkyl, hydroxy(C₁-C₃₀)alkyl, hydroxy(C₂-C₃₀)alkenyl,hydroxyphenyl(C₁-C₃₀)alkyl or hydroxynaphthyl(C₁-C₃₀)alkyl;

-   X⁶,X⁷,X⁸,X⁹,X¹⁰ for each occurrence is independently selected from    the group consisting of H, (C₁-C₆)alkyl, OH, OR⁴, NO₂, CN, and halo;-   R⁴ is (C₁-C₃₀)alkyl, (C₂-C₃₀)alkenyl, phenyl(C₁-C₃₀)alkyl,    naphthyl(C₁-C₃₀)alkyl, hydroxy(C₁-C₃₀)alkyl, hydroxy(C₂-C₃₀)alkenyl,    hydroxyphenyl(C₁-C₃₀)alkyl or hydroxynaphthyl(C₁-C₃₀)alkyl;-   e is, independently for each occurrence, an integer from 1 to 4    inclusive;-   m is, independently for each occurrence, an integer from 5 to 24    inclusive;-   s is, independently for each occurrence, an integer from 5 to 10 or    from 12 to 20 inclusive;-   n is, independently for each occurrence, an integer from 1 to 5,    inclusive;-   each of R¹⁰ and R¹¹ is, independently for each occurrence, H,    (C₁-C₃₀)alkyl, (C₁-C₃₀)acyl, (C₁-C₃₀)alkylsulfonyl, —C((NH)(NH₂)) or

and

-   R¹² and R¹³ each is, independently for each occurrence,    (C₁-C₃₀)alkyl;-   provided that:-   when A⁷ is Ura, Paa or Pta, then R² and R³ are deleted;-   when R¹⁰ is (C₁-C₃₀)acyl, (C₁-C₃₀)alkylsulfonyl, —C((NH)(NH₂)) or

then R¹¹ is H or (C₁-C₃₀)alkyl;

-   (i) at least one amino acid of a compound of formula (I) is not the    same as the native sequence of hGLP-1(7-38 or -39)NH₂ or hGLP-1(7-38    or -39)OH;-   (ii) a compound of formula (I) is not an analogue of hGLP-1(7-38 or    -39)NH₂ or hGLP-1(7-38, or -39)OH wherein a single position has been    substituted by Ala;-   (iii) a compound of formula (I) is not (Arg^(26,34),    Lys³⁸)hGLP-1(7-38)-E, (Lys²⁶(N^(ε)-alkanoyl))hGLP-1(7-38)-E,    (Lys³⁴(N^(ε)-alkanoyl))hGLP-1(7-38)-E,    (Lys^(26,34)-bis(N^(ε)-alkanoyl))hGLP-1(7-38)-E, (Arg²⁶,    Lys³⁴(N^(ε)-alkanoyl))hGLP-1(8-38)-E, (Arg^(26,34),    Lys³⁶(N^(ε)-alkanoyl))hGLP-1(7-38)-E or (Arg^(26,34),    Lys³⁸(N^(ε)-alkanoyl))hGLP-1(7-38)-E, wherein E is —OH or —NH₂;-   (iv) a compound of formula (I) is not Z¹-hGLP-1(7-38)-OH,    Z¹-hGLP-1(7-38)-NH₂; wherein Z¹ is selected from the group    consisting of:

(a) (Arg²⁶), (Arg³⁴), (Arg^(26,34)), (Lys³⁶), (Arg²⁶, Lys³⁶), (Arg³⁴,Lys³⁶), (D-Lys³⁶), (Arg³⁶), (D-Arg³⁶), (Arg^(26,34), Lys³⁶) or(Arg^(26,36), Lys³⁴);

(b) (Asp²¹);

(c) at least one of (Aib⁸), (D-Ala⁸) and (Asp⁹); and

(d) (Tyr⁷), (N-acyl-His⁷), (N-alkyl-His⁷), (N-acyl-D-His⁷) or(N-alkyl-D-His⁷); and

-   (v) a compound of formula (I) is not a combination of any two of the    substitutions listed in groups (a) to (d); or a pharmaceutically    acceptable salt thereof.

A preferred group of compounds of the immediately foregoing compound iswhere A¹¹ is Thr; A¹³ is Thr; A¹⁵ is Asp; A¹⁷ is Ser; A¹⁸ is Ser or Lys;A²¹ is Glu; A²³ is Gln or Glu; A²⁷ is Glu, Leu, Aib or Lys; and A³¹ isTrp, Phe, 1Nal or 2Nal; or a pharmaceutically acceptable salt thereof.

A preferred group of compounds of the immediately foregoing group ofcompounds is where A⁹ is Glu, N-Me-Glu or N-Me-Asp; A¹² is Phe, Acc,1Nal, 2Nal, or Aic; A¹⁶ is Val, Acc or Aib; A¹⁹ is Tyr, 1Nal or 2Nal;A²⁰ is Leu, Acc or Cha; A²⁴ is Ala, Aib or Acc; A²⁵ is Ala, Aib, Acc,Lys, Arg, hArg, Orn, HN—CH((CH₂)_(n)—N(R¹⁰R¹¹))—C(O) orHN—CH((CH₂)_(e)—X³)—C(O); A²⁸ is Phe, 1Nal or 2Nal; A²⁹ is Ile or Acc;A³⁰ is Ala or Aib; A³² is Leu, Acc or Cha; and A³³ is Val, Lys or Acc;or a pharmaceutically acceptable salt thereof.

A preferred group of compounds of the immediately foregoing group ofcompounds is where A⁸ is Ala, Gly, Ser, D-Ala, Aib, A6c, A5c, N-Me-Ala,N-Me-D-Ala or N-Me-Gly; A¹⁰ is Gly; A¹² is Phe, 1Nal, 2Nal, A6c or A5c;A¹⁶ is Val, A6c or A5c; A²⁰ is Leu, A6c, A5c or Cha; A²² is Gly, β-Ala,Glu or Aib; A²⁴ is Ala or Aib; A²⁹ is Ile, A6c or A5c; A³² is Leu, A6c,A5c or Cha; A³³ is Val, Lys, A6c or A5c; A³⁵ is Aib, β-Ala, Ado, A6c,A5c, D-Arg or Acc; A³⁷ is Gly, Aib, β-Ala, D-Ala, Pro, Asp, Aun orD-Asp; A³⁸ is D- or L-His, Asn, Ser, Apc, Act, Gly, β-Ala or Gaba; andA³⁹ is Ser, Thr or Aib; or a pharmaceutically acceptable salt thereof.

A preferred group of compounds of the immediately foregoing group ofcompounds is where X⁴ for each occurrence is —C(O)—; and R¹ is OH orNH₂; or a pharmaceutically acceptable salt thereof.

A preferred group of compounds of the immediately foregoing group ofcompounds or a pharmaceutically acceptable salt thereof is where R² is Hand R³ is (C₁-C₃₀)alkyl, (C₂-C₃₀)alkenyl, (C₁-C₃₀)acyl,(C₁-C₃₀)alkylsulfonyl,

A preferred group of the compounds of the immediately foregoing group ofcompounds or a pharmaceutically acceptable salt thereof, is where R¹⁰ is(C₁-C₃₀)acyl, (C₁-C₃₀)alkylsulfonyl or

A preferred group of the compounds of the immediately foregoing group ofcompounds or a pharmaceutically acceptable salt thereof, is where R¹⁰ is(C₄-C₂₀)acyl, (C₄-C₂₀)alkylsulfonyl or

A preferred compound of the formula (I) is where A⁸ is Ala, D-Ala, Aib,A6c, A5c, N-Me-Ala, N-Me-D-Ala or N-Me-Gly; A¹⁰ is Gly; A¹² is Phe,1Nal, 2Nal, A6c or A5c; A¹⁶ is Val, A6c or A5c; A²⁰ is Leu, A6c, A5c orCha; A²² is Gly, β-Ala, Glu or Aib; A²⁴ is Ala or Aib; A²⁹ is Ile, A6cor A5c; A³² is Leu, A6c, A5c or Cha; A³³ is Val, Lys, A6c or A5c; A³⁵ isAib, β-Ala, Ado, A6c, A5c or D-Arg; and A³⁷ is Gly, Aib, β-Ala, D-Ala,Pro or D-Asp; A³⁸ is D- or L-His, Asn, Ser, Gly, β-Ala or Gaba; and A³⁹is Ser, or deleted; X⁴ for each occurrence is —C(O)—; e for eachoccurrence is independently 1 or 2; R¹ is OH or NH₂; R¹⁰ is(C₁-C₃₀)acyl, (C₁-C₃₀)alkylsulfonyl or

and R¹¹ is H; or a pharmaceutically acceptable salt thereof.

More preferred of the immediately foregoing compounds is where R¹⁰ is(C₄-C₂₀)acyl, (C₄-C₂₀)alkylsulfonyl or

or a pharmaceutically acceptable salt thereof.

A more preferred compound of formula (I) is where said compound is thepharmaceutically acceptable salt thereof.

More preferred of the immediately foregoing group of compounds is acompound of the formula:

-   (Aib^(8,35), Arg^(26,34), Phe³¹, Pro³⁷,    Ser^(38,39))hGLP-1(7-39)-NH₂; (SEQ ID NO:1)-   (Aib^(8,35,37), Arg^(26,34), Phe³¹, Asn³⁸)hGLP-1(7-38)-NH₂; (SEQ ID    NO:2)-   (Aib^(8,35,37), Arg^(26,34), Phe³¹, Ser³⁸)hGLP-1(7-38) NH₂; (SEQ ID    NO:3)-   (Aib^(8,35,37), Gaba³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:4)-   (Aib^(8,35,37), Arg^(26,34), Phe³¹, His³⁸)hGLP-1(7-38) NH₂; (SEQ ID    NO:5)-   (Aib^(8,35), Arg^(26,34), Phe³¹, β-Ala³⁷, His³⁸)hGLP-1(7-38) NH₂;    (SEQ ID NO:6)-   (Aib^(8,35,37), Arg^(26,34), D-His³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:7)-   (Aib^(8,35,37), β-Ala³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:8)-   (Aib^(8,35), Arg^(26,34), β-Ala³⁷, His³⁸)hGLP-1(7-38) NH₂; (SEQ ID    NO:9)-   (Aib^(8,35,37), Arg^(26,34), Phe³¹, Gly³⁸)hGLP-1(7-38) NH₂; (SEQ ID    NO:10)-   (Aib^(8,35,37), Arg^(26,34), Gly³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:11)-   (Aib^(8,35,37), Arg^(26,34), β-Ala³⁸)hGLP-1(7-38) NH₂; (SEQ ID    NO:12)-   (Aib^(8,35,37), Arg^(26,34), Gaba³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:13)-   (Aib^(8,35,37), Arg³⁴, Phe³¹, His³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:14)-   (Aib^(8,35,37), Arg^(26,34), His³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:15)-   (Aib^(8,35,37), Arg^(26,34), Phe³¹, Gaba³⁸)hGLP-1(7-38) NH₂; (SEQ ID    NO:16)-   (Aib^(8,35,37), Arg^(26,34), Phe³¹, Ava³⁸)hGLP-1(7-38) NH₂; (SEQ ID    NO:17)-   (Aib^(8,35,37), Arg^(26,34), Ava³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:18)-   (Aib^(8,35,37), Arg³⁴, Phe³¹, D-His³⁸)hGLP-1(7-38) NH₂; (SEQ ID    NO:19)-   (Aib^(8,35,37), Arg³⁴, Phe³¹, Gly³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:20)-   (Aib^(8,35,37), Gly³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:21)-   (Aib^(8,35,37), Arg^(26,34), Phe³¹, D-His³⁸)hGLP-1(7-38) NH₂; (SEQ    ID NO:22)-   (Aib^(8,35), Arg^(26,34), Phe³¹, β-Ala³⁷, D-His³⁸)hGLP-1(7-38) NH₂;    (SEQ ID NO:23)-   (Aib^(8,35,37), Arg^(26,34), Phe³¹, β-Ala³⁸)hGLP-1(7-38) NH₂; (SEQ    ID NO:24)-   (Aib^(8,35), Arg^(26,34), Phe³¹, β-Ala^(37,38))hGLP-1(7-38) NH₂;    (SEQ ID NO:25)-   (Aib^(8,35,37), Arg³⁴, Phe³¹, β-Ala³⁸)hGLP-1(7-38) NH₂; (SEQ ID    NO:26) or-   (Aib^(8,35,37), Arg³⁴, Phe³¹, Gaba³⁸)hGLP-1(7-38) NH₂; (SEQ ID    NO:27)-   or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention is directed to a compoundaccording to formula (II),R⁷-A⁸-A⁹-A¹⁰-A¹¹-A¹²-A¹³-A¹⁴-A¹⁵-A¹⁶-A¹⁷-A¹⁸-A¹⁹-A²⁰-A²¹-A²²-A²³-A²⁴-A²⁵-A²⁶-A²⁷-A²⁸-A²⁹-A³⁰-A³¹-A³²-A³³-A³⁴-A³⁵-A³⁶-A³⁷-A³⁸-A³⁹-R¹,  (II)wherein

-   A⁸ is Ala, β-Ala, Gly, Ser, D-Ala, Aib, Acc, N-Me-Ala, N-Me-D-Ala or    N-Me-Gly;-   A⁹ is Glu, N-Me-Glu, N-Me-Asp or Asp;-   A¹⁰ is Gly, Acc, β-Ala or Aib;-   A¹¹ is Thr or Ser;-   A¹² is Phe, Acc, Aic, Aib, 2-Pal, 3-Pal, 4-Pal, 1Nal, 2Nal, Cha, Trp    or (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe;-   A¹³ is Thr or Ser;-   A¹⁴ is Ser or Aib;-   A¹⁵ is Asp or Glu;-   A¹⁶ is Val, Acc, Aib, Leu, Ile, Tle, Nle, Abu, Ala or Cha;-   A¹⁷ is Ser, Aib or Thr;-   A¹⁸ is Ser, Lys or Thr;-   A¹⁹ is Tyr, Cha, Phe, 2-Pal, 3-Pal, 4-Pal, 1Nal, 2Nal, Acc or    (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe;-   A²⁰ is Leu, Acc, Aib, Nle, Ile, Cha, Tle, Val, Phe or    (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe;-   A²¹ is Glu or Asp;-   A²² is Gly, Acc, β-Ala, Glu or Aib;-   A²³ is Gln, Asp, Asn or Glu;-   A²⁴ is Ala, Aib, Val, Abu, Tle or Acc;-   A²⁵ is Ala, Aib, Val, Abu, Tle, Acc, Lys, Arg, hArg, Orn,    HN—CH((CH₂)_(n)—N(R¹⁰R¹¹))—C(O) or HN—CH((CH₂)_(e)—X³)—C(O);-   A²⁶ is Lys, Arg, hArg, Orn, Lys(N^(ε)-decanoyl)),    HN—CH((CH₂)_(n)—N(R¹⁰R¹¹))—C(O) or HN—CH((CH₂)_(e)—X³)—C(O);-   A²⁷ is Glu Asp, Leu, Aib or Lys;-   A²⁸ is Phe, 2-Pal, 3-Pal, 4-Pal, 1Nal, 2Nal, (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe,    Aic, Acc, Aib, Cha or Trp;-   A²⁹ is Ile, Acc, Aib, Leu, Nle, Cha, Tle, Val, Abu, Ala or Phe;-   A³⁰ is Ala, Aib or Acc;-   A³¹ is Trp, 2-Pal, 3-Pal, 4-Pal, 1Nal, 2Nal, Phe, Acc, Aib,    (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe or Cha;-   A³² is Leu, Acc, Aib, Nle, Ile, Cha, Tle, Phe, (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe    or Ala;-   A³³ is Val, Acc, Aib, Leu, Ile, Tle, Nle, Cha, Ala, Phe, Abu, Lys or    (X⁶,X⁷,X⁸,X⁹,X¹⁰)Phe;-   A³⁴ is Lys, Arg, hArg, Orn, HN—CH((CH₂)_(n)—N(R¹⁰R¹¹))—C(O) or    HN—CH((CH₂)_(e)—X³)—C(O);-   A³⁵ is β-Ala, D-Ala, Gaba, Ava, HN—(CH₂)_(m)—C(O), Aib, Acc, D-Arg,    a D-amino acid or deleted;-   A³⁶ is L- or D-Arg, D- or L-Lys, or Lys(N^(ε)-decanoyl) or    Lys(N^(ε)-dodecanoyl) or D- or L-hArg, D- or L-Orn or    HN—CH((CH₂)_(n)—N(R¹⁰R¹¹))—C(O), HN—CH((CH₂)_(e)—X³)—C(O), or    deleted;-   A³⁷ is Gly, β-Ala, Gaba, Aib, Acc, Act, Apc, Aun, Ava, Pro, Dhp,    Dmt, Pip, L- or D-Arg, L- or D-Asp or Glu, Lys(N^(ε)-decanoyl),    Lys(N^(ε)-dodecanoyl), Lys(N^(ε)-octanoyl),    Lys(N^(ε)-tetradecanoyl), Ser(O-decanoyl), or deleted;-   A³⁸ is D- or L-His, L- or D-Ala, Asn, Gln, Ser, Thr, Acc, Ado, Aib,    Apc, Act, Arg, Ava, Gly, β-Ala, Gaba, HN—(CH₂)_(m)—C(O), or deleted;-   A³⁹ is D- or L-His, L- or D-Ala, Asn, Gln, Ser, Thr, Acc, Ado, Aib,    Apc, Act, Arg, Aun, Gly, β-Ala, Gaba, Lys(N^(ε)-octanoyl),    HN—(CH₂)_(m)—C(O), or deleted;-   R¹ is OH, NH₂; (C₁-C₃₀)alkoxy, or NH—X²—CH₂—Z⁰, wherein X² is a    (C₀-C₂₀)hydrocarbon moiety and Z⁰ is H, OH, CO₂H or CONH₂;

or —C(O)—NHR¹², wherein X⁴ is, independently for each occurrence,—C(O)—, —NH—C(O)— or —CH₂—, and wherein f is, independently for eachoccurrence, an integer from 1 to 29 inclusive;

-   X⁶,X⁷,X⁸,X⁹,X¹⁰ for each occurrence is independently selected from    the group consisting of H, (C₁-C₆)alkyl, OH, OR⁴, NO₂, CN, and halo;-   R⁴ is (C₁-C₃₀)alkyl, (C₂-C₃₀)alkenyl, phenyl(C₁-C₃₀)alkyl,    naphthyl(C₁-C₃₀)alkyl, hydroxy(C₁-C₃₀)alkyl, hydroxy(C₂-C₃₀)alkenyl,    hydroxyphenyl(C₁-C₃₀)alkyl or hydroxynaphthyl(C₁-C₃₀)alkyl;-   Z¹,Z²,Z³,Z⁴,Z⁵ for each occurrence is independently selected from    the group consisting of H, (C₁-C₆)alkyl, OH, OR⁴, NO₂, CN, and halo;    Z¹ and Z² can join together to form a ring system;-   e is, independently for each occurrence, an integer from 1 to 4    inclusive;-   m is, independently for each occurrence, an integer from 5 to 24    inclusive;-   n is, independently for each occurrence, an integer from 1 to 5,    inclusive;-   t is, independently for each occurrence, an integer from 0 to 4,    inclusive;-   each of R¹⁰ and R¹¹ is, independently for each occurrence, H,    (C₁-C₃₀)alkyl, (C₁-C₃₀)acyl, (C₁-C₃₀)alkylsulfonyl, —C((NH)(NH₂)) or

and

-   R¹² and R¹³ each is, independently for each occurrence,    (C₁-C₃₀)alkyl;-   provided that:-   R⁷ is not C(O)X¹¹, wherein X¹¹ is phenyl(C₁-C₃₀)alkyl,    naphthyl(C₁-C₃₀)alkyl, hydroxy(C₁-C₃₀)alkyl, hydroxy(C₂-C₃₀)alkenyl,    hydroxyphenyl(C₁-C₃₀)alkyl or hydroxynaphthyl(C₁-C₃₀)alkyl;    when R¹⁰ is (C₁-C₃₀)acyl, (C₁-C₃₀)alkylsulfonyl, —C((NH)(NH₂)) or

then R¹¹ is H or (C₁-C₃₀)alkyl;

-   or a pharmaceutically acceptable salt thereof.

A preferred group of compounds of the immediately foregoing compound iswhere A¹¹ is Thr; A¹³ is Thr; A¹⁵ is Asp; A¹⁷ is Ser; A¹⁸ is Ser or Lys;A²¹ is Glu; A²³ is Gln or Glu; A²⁷ is Glu, Leu, Aib or Lys; and A³¹ isTrp, Phe, 1Nal or 2Nal; or a pharmaceutically acceptable salt thereof.

A preferred group of compounds of the immediately foregoing group ofcompounds is where A⁷ is 4-imidazol-carbonyl, 4-nitrophenyl-acetyl,3-chloro-4-hydroxyphenyl-acetyl, 4-hydroxyphenyl-acetyl,3-(4-aminophenyl)-propionyl, 3-(4-nitrophenyl)-propionyl,3-(3,4-difluorophenyl)-propionyl, 3-fluoro-4-hydroxyphenyl-acetyl or4-aminophenyl-acetyl; A⁹ is Glu, N-Me-Glu or N-Me-Asp; A¹² is Phe, Acc,1Nal, 2Nal or Aic; A¹⁶ is Val, Acc or Aib; A¹⁹ is Tyr, 1Nal or 2Nal; A²⁰is Leu, Acc or Cha; A²⁴ is Ala, Aib or Acc; A²⁵ is Ala, Aib, Acc, Lys,Arg, hArg, Orn, HN—CH((CH₂)_(n)—N(R¹⁰R¹¹))—C(O) orHN—CH((CH₂)_(e)—X³)—C(O); A²⁸ is Phe, 1Nal or 2Nal; A²⁹ is Ile or Acc;A³⁰ is Ala or Aib; A³² is Leu, Acc or Cha; and A³³ is Val, Lys or Acc;or a pharmaceutically acceptable salt thereof.

A preferred group of compounds of the immediately foregoing group ofcompounds is where A⁸ is Ala, Gly, Ser, D-Ala, Aib, A6c, A5c, N-Me-Ala,N-Me-D-Ala or N-Me-Gly; A¹⁰ is Gly; A¹² is Phe, 1Nal, 2Nal, A6c or A5c;A¹⁶ is Val, A6c or A5c; A²⁰ is Leu, A6c, A5c or Cha; A²² is Gly, β-Ala,Glu or Aib; A²⁴ is Ala or Aib; A²⁹ is Ile, A6c or A5c; A³² is Leu, A6c,A5c or Cha; A³³ is Val, Lys, A6c or A5c; A³⁵ is Aib, β-Ala, Ado, A6c,A5c, D-Arg, Acc or Gly; A³⁷ is Gly, Aib, β-Ala, D-Ala, Pro, Asp, Aun orD-Asp; A³⁸ is D- or L-His, Asn, Ser, Apc, Act, Gly, β-Ala or Gaba; andA³⁹ is Ser, Thr or Aib; or a pharmaceutically acceptable salt thereof.

A preferred group of compounds of the immediately foregoing group ofcompounds is where X⁴ for each occurrence is —C(O)—; and R¹ is OH orNH₂; or a pharmaceutically acceptable salt thereof.

A preferred compound of the formula (II) is where A⁸ is Ala, D-Ala, Aib,A6c, A5c, N-Me-Ala, N-Me-D-Ala or N-Me-Gly; A¹⁰ is Gly; A¹² is Phe,1Nal, 2Nal, A6c or A5c; A¹⁶ is Val, A6c or A5c; A²⁰ is Leu, A6c, A5c orCha; A²² is Gly, β-Ala, Glu or Aib; A²⁴ is Ala or Aib; A²⁹ is Ile, A6cor A5c; A³² is Leu, A6c, A5c or Cha; A³³ is Val, Lys, A6c or A5c; A³⁵ isAib, β-Ala, Ado, A6c, A5c D-Arg or Gly; and A³⁷ is Gly, Aib, β-Ala,D-Ala, Pro or D-Asp; A³⁸ is D- or L-His, Asn, Ser, Gly, β-Ala or Gaba;and A³⁹ is Ser, or deleted; X⁴ for each occurrence is —C(O)—; e for eachoccurrence is independently 1 or 2; R¹ is OH or NH₂; R¹⁰ is(C₁-C₃₀)acyl, (C₁-C₃₀)alkylsulfonyl or

and R¹¹ is H; or a pharmaceutically acceptable salt thereof.

More preferred of the immediately foregoing compounds is where R¹⁰ is(C₄-C₂₀)acyl, (C₄-C₂₀)alkylsulfonyl or

or a pharmaceutically acceptable salt thereof.

A more preferred compound of formula (II) is where said compound is ofthe formula:

-   (4Hppa⁷)GLP-1(7-36)NH₂; (SEQ ID NO:28)-   (3Hppa⁷)GLP-1(7-36)NH_(2;) (SEQ ID NO:29)-   (phenylacetyl⁷)hGLP-1(7-36)NH₂; (SEQ ID NO:30)-   ((3-fluoro-4-hydroxyphenyl-acetyl)⁷)hGLP-1(7-36)NH₂; (SEQ ID NO:31)-   ((4-imidazol-carbonyl)⁷)hGLP-1(7-36) NH₂; (SEQ ID NO:32)-   ((4-nitrophenyl-acetyl)⁷)hGLP-1(7-36) NH₂; (SEQ ID NO:33)-   ((3-chloro-4-hydroxyphenyl-acetyl)⁷)hGLP-1(7-36) NH₂; (SEQ ID NO:34)-   ((4-hydroxyphenylacetyl)⁷)hGLP-1(7-36) NH₂; (SEQ ID NO:35)-   ((4-aminophenyl-acetyl)⁷)hGLP-1(7-36) NH₂; (SEQ ID NO:36)-   ((3-(3-hydroxyphenyl)-propionyl)⁷)hGLP-1(7-36) NH₂; (SEQ ID NO:37)-   ((3-phenyl-propionyl)⁷)hGLP-1(7-36) NH₂; (SEQ ID NO:38)-   ((3-(4-aminophenyl)-propionyl)⁷)hGLP-1(7-36) NH₂; (SEQ ID NO:39)-   ((3-(4-nitrophenyl)-propionyl)⁷)hGLP-1(7-36) NH₂; (SEQ ID NO:40)-   ((3-(2-hydroxyphenyl)-propionyl)⁷)hGLP-1(7-36) NH₂; (SEQ ID NO:41)-   ((3-(3,4-difluorophenyl)-propionyl)⁷)hGLP-1(7-36) NH₂; (SEQ ID    NO:42) or-   ((3-(2,4-dihydroxyphenyl)-propionyl)⁷)hGLP-1(7-36) NH₂; (SEQ ID    NO:43)-   or a pharmaceutically acceptable salt thereof.

A more preferred compound of formula (II) is where said compound is thepharmaceutically acceptable salt thereof.

Another more preferred compound of formula (I) or (II) is each of thecompounds that are specifically enumerated hereinbelow in the Examplessection of the present disclosure, or a pharmaceutically acceptable saltthereof.

In another aspect, the present invention provides a pharmaceuticalcomposition comprising an effective amount of a compound of formula (I)or (II) as defined hereinabove or a pharmaceutically acceptable saltthereof and a pharmaceutically acceptable carrier or diluent.

In yet another aspect, the present invention provides a method ofeliciting an agonist effect from a GLP-1 receptor in a subject in needthereof which comprises administering to said subject an effectiveamount of a compound of formula (I) or (II) as defined hereinabove or apharmaceutically acceptable salt thereof.

In a further aspect, the present invention provides a method of treatinga disease selected from the group consisting of Type I diabetes, Type IIdiabetes, obesity, glucagonomas, secretory disorders of the airway,metabolic disorder, arthritis, osteoporosis, central nervous systemdisease, restenosis, neurodegenerative disease, renal failure,congestive heart failure, nephrotic syndrome, cirrhosis, pulmonaryedema, hypertension, treatment of respiratory distress (U.S. PatentApplication Publication No. 2004/0235726 A1) and disorders wherein thereduction of food intake is desired, in a subject in need thereof whichcomprises administering to said subject an effective amount of acompound of formula (I) or (II) as defined hereinabove or apharmaceutically acceptable salt thereof. A preferred method of theimmediately foregoing method is where the disease being treated is TypeI diabetes or Type II diabetes. GLP-1 analogues of the present inventionthat elicit an antagonist effect from a subject can be used for treatingthe following: hypoglycemia and malabsorption syndrome associated withgastroectomy or small bowel resection.

With the exception of the N-terminal amino acid, all abbreviations (e.g.Ala) of amino acids in this disclosure stand for the structure of—NH—CH(R)—CO—, wherein R and R′ each is, independently, hydrogen or theside chain of an amino acid (e.g., R═CH₃ and R′═H for Ala) or R and R′may be joined to form a ring system. For the N-terminal amino acid, theabbreviation stands for the structure of ═N—C(R)(R′)—CO—, wherein “═”represents the bonds to R² and R³, defined herein. R² and R³ are asdefined above, except when A⁷ is Ura, Paa or Pta, in which case R² andR³ are not present since Ura, Paa and Pta are considered here asdes-amino amino acids.

The application employs the following commonly understood abbreviations:

Abu α-aminobutyric acid Acc 1-amino-1-cyclo(C₃—C₉)alkyl carboxylic acidA3c 1-amino-1-cyclopropanecarboxylic acid A4c1-amino-1-cyclobutanecarboxylic acid A5c1-amino-1-cyclopentanecarboxylic acid A6c1-amino-1-cyclohexanecarboxylic acid Act4-amino-4-carboxytetrahydropyran Ado 12-aminododecanoic acid Aec4-(2-aminoethyl)-1-carboxymethyl-piperazine

Aib α-aminoisobutyric acid Aic 2-aminoindan-2-carboxylic acid Ala or Aalanine β-Ala beta-alanine Amp 4-amino-phenylalanine; Apc4-amino-4-carboxypiperidine: Arg or R arginine hArg homoarginine Asn orN asparagine Asp or D aspartic acid Aun 11-aminoundecanoic acid Ava5-aminovaleric acid Cha β-cyclohexylalanine Dhp 3,4-dehydroproline Dmt5,5-dimethylthiazolidine-4-carboxylic acid Gaba γ-aminobutyric acid Glnor Q glutamine Glu or E glutamic acid Gly or G glycine His or Hhistidine 4Hppa 3-(4-hydroxyphenyl)propionic acid 3Hppa3-(3-hydroxyphenyl)propionic acid 3Hyp trans-3-hydroxy-L-proline (i.e.,(2S, 3S)-3-hydroxypyrrolidine-2-carboxylic acid) 4Hyp 4-hydroxyproline(i.e., (2S, 4R)-4-hydroxypyrrolidine-2-carboxylic acid) Ile or Iisoleucine Leu or L leucine Lys or K lysine 1Nal β-(1-naphthyl)alanine2Nal β-(2-naphthyl)alanine Nle norleucine N-Me-Ala N-methyl-alanine;N-Me-Asp N-methyl-aspartic acid N-Me-Glu N-methyl-glutamic acid;N-Me-Gly N-methyl-glycine; Nva norvaline Orn ornithine Paatrans-3-(3-pyridyl) acrylic acid; 2Pal β-(2-pyridinyl)alanine 3Palβ-(3-pyridinyl)alanine 4Pal β-(4-pyridinyl)alanine Phe or Fphenylalanine (3,4,5F)Phe 3,4,5-trifluorophenylalanine (2,3,4,5,6)Phe2,3,4,5,6-pentafluorophenylalanine Pip pipecolic acid Pro or P prolinePta (4-pyridylthio) acetic acid; Ser or S serine Thr or T threonine Tletert-leucine Tma-His N,N-tetramethylamidino-histidine; Trp or Wtryptophan Tyr or Y tyrosine Ura urocanic acid. Val or V valineCertain other abbreviations used herein are defined as follows:

2BrZ 2-bromobenzyloxycarbonyl 2ClZ 2-chlorobenzyloxycarbonyl Boc:tert-butyloxycarbonyl Bzl: benzyl DCM: dichloromethane DIC:N,N-diisopropylcarbodiimide DIEA: diisopropylethyl amine Dmab:4-{N-(1-(4,4-dimethyl-2,6-dioxocyclohexylidene)- 3-methylbutyl)-amino}benzyl DMAP: 4-(dimethylamino)pyridine DMF dimethylformamide DNP:2,4-dinitrophenyl Fm formyl Fmoc: 9-Fluorenylmethyloxycarbonyl HBTU:2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphatecHex cyclohexyl HF hydrogen fluoride, HOAT:O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate HOBt: 1-hydroxy-benzotriazole Mmt: 4-methoxytritylNMP: N-methylpyrrolidone OcHex O-cyclohexyl PAM4-hydroxymethylphenylacetamidomethyl resin resin Pbf:2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl tBu: tert-butyl TIS:triisopropylsilane TOS: tosyl trt trityl TFA: trifluoro acetic acidTFFH: tetramethylfluoroforamidinium hexafluorophosphate Xan xanthyl Z:benzyloxycarbonyl

In the above formula, hydroxyalkyl, hydroxyphenylalkyl, andhydroxynaphthylalkyl may contain 1-4 hydroxy substituents. COX⁵ standsfor —C═O·X⁵. Examples of —C═O·X⁵ include, but are not limited to, acetyland phenylpropionyl.

What is meant by Lys(N^(ε)-alkanoyl) is represented by the followingstructure:

What is meant by Lys(N^(ε)-alkylsulfonyl) is represented by thefollowing structure:

What is meant by Lys(N^(ε)-(2-(4-alkyl-1-piperazine)acetyl)) isrepresented by the following structure:

What is meant by Asp(1-(4-alkyl-piperazine)) is represented by thefollowing structure:

What is meant by Asp(1-alkylamino) is represented by the followingstructure:

What is meant by Lys(N^(ε)-Aec-alkanoyl) is represented by thestructure:

The variable n in the foregoing structures is 1-30.Lys(N^(ε)-Aec-alkanoyl) is represented by the structure:

The term “halo” encompasses fluoro, chloro, bromo and iodo.

The term “(C₁-C₃₀)hydrocarbon moiety” encompasses alkyl, alkenyl andalkynyl, and in the case of alkenyl and alkynyl there are C₂-C₃₀.

A peptide of this invention is also denoted herein by another format,e.g., (A5c⁸)hGLP-1(7-36)NH₂; with the substituted amino acids from thenatural sequence placed between the first set of parentheses (e.g., A5c⁸for Ala⁸ in hGLP-1). The abbreviation GLP-1 means glucagon-likepeptide-1; hGLP-1 means human glucagon-like peptide-1. The numbersbetween the parentheses refer to the number of amino acids present inthe peptide (e.g., hGLP-1(7-36) is amino acids 7 through 36 of thepeptide sequence for human GLP-1). The sequence for hGLP-1(7-37) islisted in Mojsov, S., Int. J. Peptide Protein Res,. 40, 1992, pp.333-342. The designation “NH₂” in hGLP-1(7-36)NH₂ indicates that theC-terminus of the peptide is amidated. hGLP-1(7-36) means that theC-terminus is the free acid. In hGLP-1(7-38), residues in positions 37and 38 are Gly and Arg, respectively.

DETAILED DESCRIPTION

The peptides of this invention can be prepared by standard solid phasepeptide synthesis. See, e.g., Stewart, J. M., et al., Solid PhaseSynthesis (Pierce Chemical Co., 2d ed. 1984). The substituents R² and R³of the above generic formula may be attached to the free amine of theN-terminal amino acid by standard methods known in the art. For example,alkyl groups, e.g., (C₁-C₃₀)alkyl, may be attached using reductivealkylation. Hydroxyalkyl groups, e.g., (C₁-C₃₀)hydroxyalkyl, may also beattached using reductive alkylation wherein the free hydroxy group isprotected with a t-butyl ester. Acyl groups, e.g., COE¹, may be attachedby coupling the free acid, e.g., E¹COOH, to the free amine of theN-terminal amino acid by mixing the completed resin with 3 molarequivalents of both the free acid and diisopropylcarbodiimide inmethylene chloride for one hour. If the free acid contains a freehydroxy group, e.g., 3-fluoro-4-hydroxyphenylacetic acid, then thecoupling should be performed with an additional 3 molar equivalents ofHOBT.

When R¹ is NH—X²—CH₂—CONH₂; (i.e., Z⁰═CONH₂), the synthesis of thepeptide starts with BocHN—X²—CH₂—COOH which is coupled to the MBHAresin. If R¹ is NH—X²—CH₂—COOH, (i.e., Z⁰═COOH) the synthesis of thepeptide starts with Boc-HN—X²—CH₂—COOH which is coupled to PAM resin.For this particular step, 4 molar equivalents of Boc-HN—X²—COOH, HBTUand HOBt and 10 molar equivalents of DIEA are used. The coupling time isabout 8 hours.

In the synthesis of a GLP-1 analogue of this invention containing A5c,A6c, and/or Aib, the coupling time is 2 hrs for these residues and theresidue immediately following them.

The substituents R² and R³ of the above generic formula can be attachedto the free amine of the N-terminal amino acid by standard methods knownin the art. For example, alkyl groups, e.g., (C₁-C₃₀)alkyl, can beattached using reductive alkylation. Hydroxyalkyl groups, e.g.,(C₁-C₃₀)hydroxyalkyl, can also be attached using reductive alkylationwherein the free hydroxy group is protected with a t-butyl ester. Acylgroups, e.g., COX⁵, can be attached by coupling the free acid, e.g.,X⁵COOH, to the free amine of the N-terminal amino acid by mixing thecompleted resin with 3 molar equivalents of both the free acid anddiisopropylcarbodiimide in methylene chloride for about one hour. If thefree acid contains a free hydroxy group, e.g.,3-fluoro-4-hydroxyphenylacetic acid, then the coupling should beperformed with an additional 3 molar equivalents of HOBT.

The peptides of this invention can be provided in the form ofpharmaceutically acceptable salts. Examples of such salts include, butare not limited to, those formed with organic acids (e.g., acetic,lactic, maleic, citric, malic, ascorbic, succinic, benzoic,methanesulfonic, toluenesulfonic, or pamoic acid), inorganic acids(e.g., hydrochloric acid, sulfuric acid, or phosphoric acid), andpolymeric acids (e.g., tannic acid, carboxymethyl cellulose, polylactic,polyglycolic, or copolymers of polylactic-glycolic acids). A typicalmethod of making a salt of a peptide of the present invention is wellknown in the art and can be accomplished by standard methods of saltexchange. Accordingly, the TFA salt of a peptide of the presentinvention (the TFA salt results from the purification of the peptide byusing preparative HPLC, eluting with TFA containing buffer solutions)can be converted into another salt, such as an acetate salt bydissolving the peptide in a small amount of 0.25 N acetic acid aqueoussolution. The resulting solution is applied to a semi-prep HPLC column(Zorbax, 300 SB, C-8). The column is eluted with (1) 0.1N ammoniumacetate aqueous solution for 0.5 hrs, (2) 0.25N acetic acid aqueoussolution for 0.5 hrs and (3) a linear gradient (20% to 100% of solutionB over 30 min) at a flow rate of 4 ml/min (solution A is 0.25N aceticacid aqueous solution; solution B is 0.25N acetic acid inacetonitrile/water, 80:20). The fractions containing the peptide arecollected and lyophilized to dryness.

As is well known to those skilled in the art, the known and potentialuses of GLP-1 is varied and multitudinous (See, Todd, J. F., et al.,Clinical Science, 1998, 95, pp. 325-329; and Todd, J. F. et al.,European Journal of Clinical Investigation, 1997, 27, pp.533-536). Thus,the administration of the compounds of this invention for purposes ofeliciting an agonist effect can have the same effects and uses as GLP-1itself. These varied uses of GLP-1 may be summarized as follows,treatment of: Type I diabetes, Type II diabetes, obesity, glucagonomas,secretory disorders of the airway, metabolic disorder, arthritis,osteoporosis, central nervous system diseases, restenosis,neurodegenerative diseases, renal failure, congestive heart failure,nephrotic syndrome, cirrhosis, pulmonary edema, hypertension, treatmentof respiratory distress (U.S. Patent Application Publication No.2004/0235726 A1), and disorders wherein the reduction of food intake isdesired. GLP-1 analogues of the present invention that elicit anantagonist effect from a subject can be used for treating the following:hypoglycemia and malabsorption syndrome associated with gastroectomy orsmall bowel resection.

Accordingly, the present invention includes within its scopepharmaceutical compositions comprising, as an active ingredient, atleast one of the compounds of formula (I) or (II) in association with apharmaceutically acceptable carrier.

The dosage of active ingredient in the compositions of this inventionmay be varied; however, it is necessary that the amount of the activeingredient be such that a suitable dosage form is obtained. The selecteddosage depends upon the desired therapeutic effect, on the route ofadministration, and on the duration of the treatment. In general, aneffective dosage for the activities of this invention is in the range of1×10⁻⁷ to 200 mg/kg/day, preferably 1×10⁻⁴ to 100 mg/kg/day, which canbe administered as a single dose or divided into multiple doses.

The compounds of this invention can be administered by oral, parenteral(e.g., intramuscular, intraperitoneal, intravenous or subcutaneousinjection, or implant), nasal, vaginal, rectal, sublingual or topicalroutes of administration and can be formulated with pharmaceuticallyacceptable carriers to provide dosage forms appropriate for each routeof administration.

Solid dosage forms for oral administration include capsules, tablets,pills, powders and granules. In such solid dosage forms, the activecompound is admixed with at least one inert pharmaceutically acceptablecarrier such as sucrose, lactose, or starch. Such dosage forms can alsocomprise, as is normal practice, additional substances other than suchinert diluents, e.g., lubricating agents such as magnesium stearate. Inthe case of capsules, tablets and pills, the dosage forms may alsocomprise buffering agents. Tablets and pills can additionally beprepared with enteric coatings.

Liquid dosage forms for oral administration include, without limitation,pharmaceutically acceptable emulsions, solutions, suspensions, syrups,elixirs, and the like, containing inert diluents commonly used in theart, such as water. Besides such inert diluents, compositions can alsoinclude adjuvants, such as wetting agents, emulsifying and suspendingagents, and sweetening, flavoring and perfuming agents.

Preparations according to this invention for parenteral administrationinclude, without limitation, sterile aqueous or non-aqueous solutions,suspensions, emulsions, and the like. Examples of non-aqueous solventsor vehicles are propylene glycol, polyethylene glycol, vegetable oils,such as olive oil and corn oil, gelatin, and injectable organic esterssuch as ethyl oleate. Such dosage forms may also contain adjuvants suchas preserving, wetting, emulsifying, and dispersing agents. They may besterilized by, for example, filtration through a bacteria-retainingfilter, by incorporating sterilizing agents into the compositions, byirradiating the compositions, or by heating the compositions. They canalso be manufactured in the form of sterile solid compositions which canbe dissolved in sterile water, or some other sterile injectable mediumimmediately before use.

Compositions for rectal or vaginal administration are preferablysuppositories which may contain, in addition to the active substance,excipients such as coca butter or a suppository wax.

Compositions for nasal or sublingual administration are also preparedwith standard excipients well known in the art.

Further, a compound of this invention can be administered in a sustainedrelease composition such as those described in the following patents andpatent applications. U.S. Pat. No. 5,672,659 teaches sustained releasecompositions comprising a bioactive agent and a polyester. U.S. Pat. No.5,595,760 teaches sustained release compositions comprising a bioactiveagent in a gelable form. U.S. Pat. No. 5,821,221, teaches polymericsustained release compositions comprising a bioactive agent andchitosan. U.S. Pat. No.5,916,883 teaches sustained release compositionscomprising a bioactive agent and cyclodextrin. PCT PublicationWO99/38536 teaches absorbable sustained release compositions of abioactive agent. PCT Publication WO00/04916 teaches a process for makingmicroparticles comprising a therapeutic agent such as a peptide in anoil-in-water process. PCT Publication WO00/09166 teaches complexescomprising a therapeutic agent such as a peptide and a phosphorylatedpolymer. PCT Publication WO00/25826 teaches complexes comprising atherapeutic agent such as a peptide and a polymer bearing anon-polymerizable lactone.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Also, all publications, patentapplications, patents and other references mentioned herein are herebyincorporated by reference, each in its entirety.

The following examples describe synthetic methods for making a peptideof this invention, which methods are well-known to those skilled in theart. Other methods are also known to those skilled in the art. Theexamples are provided for the purpose of illustration and are not meantto limit the scope of the present invention in any manner.

Boc-βAla-OH, Boc-D-Arg(Tos)-OH and Boc-D-Asp(OcHex) were purchased fromNova Biochem, San Diego, Calif. Boc-Aun-OH was purchased from Bachem,King of Prussia, Pa. Boc-Ava-OH and Boc-Ado-OH were purchased fromChem-Impex International, Wood Dale, Ill. Boc-Nal-OH was purchased fromSynthetech, Inc. Albany, Oreg.

EXAMPLE 1 ((3-fluoro-4-hydroxyphenyl-acetyl)⁷)hGLP-1(7-36)NH₂(SEQ IDNO:31)

The title peptide, also referred to herein as ((3F,4HO)-phenylacetyl⁷)hGLP-1(7-36)NH₂; was synthesized on an AppliedBiosystems model 433A peptide synthesizer (Foster City, Calif.) usingFluorenylmethyloxycarbonyl (Fmoc) chemistry. A RinkAmide-4-methylbenzylhydrylamine (MBHA) resin (Novabiochem., San Diego,Calif.) with substitution of 0.66 mmol/g was used. The Fmoc amino acids(AnaSpec, San Jose, Calif.) used were Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH,Fmoc-Asp(tBu)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Glu(tBu)-OH, Fmoc-Gly-OHFmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Lys(Boc)-OH, Fmoc-Phe-OH,Fmoc-Ser(tBu)-OH, Fmco-Tyr(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Trp(Boc)-OH,and Fmoc-Val-OH. The last residue coupled to the resin was3-Fluoro-4-hydroxyphenylacetic acid (Aldrich, Milwaukee, Wis.). Thesynthesis was carried out on a 0.1 mmol scale. The Fmoc groups wereremoved by treatment with 20% piperidine in N-methylpyrrolidone (NMP)for 30 min. In each coupling step, the Fmoc amino acid (3 eq, 0.3 mmol)was first pre-activated in 2 ml solution of 0.45 M2-(1-H-benzotriazole-1-yl)-1,1,2,3-tetramethyluroniumhexafluorophosphate/1-hydroxy-benzotriazole (HBTU/HOBT) in NMP. Thisactivated amino acid ester, 1 ml of diisopropylethylamine (DIEA) and 1ml of NMP were added to the resin. The ABI 433A peptide synthesizer wasprogrammed to perform the following reaction cycle: (1) washing withNMP, (2) removing Fmoc protecting group with 20% piperidine in NMP for30 min, (3) washing with NMP, (4) coupling with pre-activated Fmoc aminoacid for 1 h. The resin was coupled successively according to thesequence of the title peptide. After the peptide chain was assembled theresin was washed completely by using N,N-dimethylformamide (DMF) anddichloromethane (DCM).

At the end of the assembly of the peptide chain, the peptide-resin wastransferred to a reaction vessel on a shaker and treated with a mixtureof TFA, H₂O and triisopropylsilane (TIS) (9.5 ml/0.85 ml/0.8 ml) for 4h. The resin was filtered off and the filtrate was poured into 200 ml ofether. The precipitate was collected by filtration and washed thoroughlywith ether. This crude product was dissolved in a mixture ofacetonitrile and aqueous acetic acid solution and purified on areverse-phase preparative HPLC system with a column (4×43 cm) of C₁₈DYNAMAX-100 A⁰ (Varian, Walnut Creek, Calif.). The column was elutedover approximately 1 hour using a linear gradient of 90% A:10% B to 50%A:50% B, where A was 0.1% TFA in water and B was 0.1% TFA inacetonitrile. The fractions were checked by analytical HPLC and thosecontaining pure product were pooled and lyophilized to dryness to give5.6 mg (1.7% yield) of a white solid. Purity was checked by using ananalytical HPLC system and found to be 95.1%. Electro-spray ionizationmass spectrometry (ESI-MS) analysis gave the molecular weight at 3312.3(in agreement with the calculated molecular weight of 3312.6).

EXAMPLE 2 (Aib^(8,35), Arg^(26,34), Phe³¹, Pro³⁷,Ser^(38,39))hGLP-1(7-39)-NH₂(SEQ ID NO:1)

The title compound was synthesized substantially according to theprocedure described for Example 1 using the appropriate protected aminoacids (AnaSpec, San Jose, Calif.). At the end of the assembly of theprotected peptide chain, an additional step was added to remove theN-terminal Fmoc-protecting group by using 20% piperidine in NMP for 30min. The peptide resin was then washed, cleaved, purified andcharacterized using the procedures described for Example 1. Yield was7.9%. Purity was 95.0%. Electro-spray ionization mass spectrometry(ESI-MS) analysis gave the molecular weight at 3629.40 (in agreementwith the calculated molecular weight of 3628.00).

The following examples can be made according to the appropriateprocedures described hereinabove:

-   Example 3 (Aib^(8,35,37), Arg^(26,34), Phe³¹,    Asn³⁸)hGLP-1(7-38)-NH₂(SEQ ID NO:2)-   Example 4 ((4-imidazol-carbonyl)⁷)hGLP-1(7-36)NH₂(SEQ ID NO:32)-   Example 5 ((3-(3-hydroxyphenyl)-propionyl)⁷)hGLP-1(7-36)NH₂(SEQ ID    NO:37)-   Example 6 ((3-phenyl-propionyl)⁷)hGLP-1(7-36)NH₂(SEQ ID NO:38)-   Example 7 ((4-nitrophenyl-acetyl)⁷)hGLP-1(7-36)NH₂(SEQ ID NO:33)-   Example 8 ((3-chloro-4-hydroxyphenyl-acetyl)⁷)hGLP-1(7-36)NH₂(SEQ ID    NO:34)-   Example 9 ((4-hydroxyphenylacetyl)⁷)hGLP-1(7-36)NH₂ (SEQ ID NO:35)-   Example 10 (Aib^(8,35,37), Arg^(26,34), Phe³¹,    Ser³⁸)hGLP-1(7-38)NH₂(SEQ ID NO:3)-   Example 11 (Aib^(8,35,37), Gaba³⁸)hGLP-1(7-38)NH₂(SEQ ID NO:4)-   Example 12 (Aib^(8,35,37), Arg^(26,34), Phe³¹,    His³⁸)hGLP-1(7-38)NH₂(SEQ ID NO:5)-   Example 13 (Aib^(8,35), Arg^(26,34), Phe³¹, β-Ala³⁷,    His³⁸)hGLP-1(7-38)NH₂(SEQ ID NO:6)-   Example 14 (Aib^(8,35,37), Arg^(26,34), D-His³⁸)hGLP-1(7-38)NH2 (SEQ    ID NO:7)-   Example 15 (Aib^(8,35,37), β-Ala³⁸)hGLP-1(7-38)NH2 (SEQ ID NO:8)-   Example 16 ((3-(4-aminophenyl)-propionyl)⁷)hGLP-1(7-36)NH₂ (SEQ ID    NO:39)-   Example 17 ((3-(4-nitrophenyl)-propionyl)⁷)hGLP-1(7-36)NH₂ (SEQ ID    NO:40)-   Example 18 ((3-(2-hydroxyphenyl)-propionyl)⁷)hGLP-1(7-36)NH₂ (SEQ ID    NO:41)-   Example 19 ((3-(3,4-difluorophenyl)-propionyl)⁷)hGLP-1(7-36)NH₂ (SEQ    ID NO:42)-   Example 20 (Aib^(8,35), Arg^(26,34), β-Ala³⁷, His³⁸)hGLP-1(7-38)NH₂    (SEQ ID NO:9)-   Example 21 (Aib^(8,35,37), Arg^(26,34), Phe³¹, Gly³⁸)hGLP-1(7-38)NH₂    (SEQ ID NO:10)-   Example 22 (Aib^(8,35,37), Arg^(26,34), Gly³⁸)hGLP-1(7-38)NH₂ (SEQ    ID NO:11)-   Example 23 (Aib^(8,35,37), Arg^(26,34), β-Ala³⁸)hGLP-1(7-38)NH₂ (SEQ    ID NO:12)-   Example 24 (Aib^(8,35,37), Arg^(26,34), Gaba³⁸)hGLP-1(7-38)NH₂ (SEQ    ID NO:13)-   Example 25 (Aib^(8,35,37), Arg³⁴, Phe³¹, His³⁸)hGLP-1(7-38)NH₂ (SEQ    ID NO:14)-   Example 26 (Aib^(8,35,37), Arg^(26,34), His³⁸)hGLP-1(7-38)NH₂ (SEQ    ID NO:15)-   Example 27 (Aib^(8,35,37), Arg^(26,34), Phe³¹,    Gaba³⁸)hGLP-1(7-38)NH₂ (SEQ ID NO:16)-   Example 28 (Aib^(8,35,37), Arg^(26,34), Phe³¹, Ava³⁸)hGLP-1(7-38)NH₂    (SEQ ID NO:17)-   Example 29 (Aib^(8,35,37), Arg^(26,34), Ava³⁸)hGLP-1(7-38)NH₂ (SEQ    ID NO:18)-   Example 30 (Aib^(8,35,37), Arg³⁴, Phe³¹, D-His³⁸)hGLP-1(7-38)NH₂    (SEQ ID NO:19)-   Example 31 (Aib^(8,35,37), Arg³⁴, Phe³¹, Gly³⁸)hGLP-1(7-38)NH₂ (SEQ    ID NO:20)-   Example 32 ((4-aminophenyl-acetyl)⁷)hGLP-1(7-36)NH₂ (SEQ ID NO:36)-   Example 33 (Aib^(8,35,37), Gly³⁸)hGLP-1(7-38)NH₂ (SEQ ID NO:21)-   Example 34 (Aib^(8,35,37), Arg^(26,34), Phe³¹,    D-His³⁸)hGLP-1(7-38)NH₂ (SEQ ID NO:22)-   Example 35 (Aib^(8,35), Arg^(26,34), Phe³¹, β-Ala³⁷,    D-His³⁸)hGLP-1(7-38)NH₂ (SEQ ID NO:23)-   Example 36 (Aib^(8,35,37), Arg^(26,34), Phe³¹,    β-Ala³⁸)hGLP-1(7-38)NH₂ (SEQ ID NO:24)-   Example 37 (Aib^(8,35), Arg^(26,34), Phe³¹,    β-Ala^(37,38))hGLP-1(7-38)NH₂ (SEQ ID NO:25)-   Example 38 (Aib^(8,35,37), Arg³⁴, Phe³¹, β-Ala³⁸)hGLP-1(7-38)NH₂    (SEQ ID NO:26)-   Example 39 (Aib^(8,35,37), Arg³⁴, Phe³¹, Gaba³⁸)hGLP-1(7-38)NH₂ (SEQ    ID NO:27)-   Example 40 ((3-(2,4-dihydroxyphenyl)-propionyl)⁷)hGLP-1(7-36)NH₂    (SEQ ID NO:43)

Physical data for a representative sampling of the compounds exemplifiedherein are given in Table 1.

TABLE 1 Example Molecular Weight Molecular Weight Purity (%) NumberCalculated MS(ES) (HPLC) 1 3312.60 3312.30 95.1 2 3628.00 3629.40 95.0 33555.94 3556.50 99.0 4 3254.59 3254.50 97.0 5 3308.68 3309.60 99.0 63292.68 3392.50 99.0 7 3323.65 3323.60 96.0 8 3329.10 3329.00 97.2 93294.65 3294.50 99.0 10 3528.91 3532.9 97.5 11 3509.95 3509.33 97.7 123578.98 3579.20 99.9 13 3564.95 3565.05 99.9 14 3618.01 3618.20 99.9 153495.92 3495.60 99.9 16 3307.69 3307.90 99.0 17 3337.68 3337.40 97.0 183308.68 3308.60 98.0 19 3328.66 3328.50 97.0 20 3603.99 3603.86 99.3 213498.89 3499.29 99.9 22 3537.92 3538.19 97.4 23 3551.95 3552.80 99.9 243565.98 3565.62 99.9 25 3550.96 3550.90 99.9 26 3618.01 3618.00 97.0 273526.94 3527.20 99.9 28 3540.97 3540.30 99.1 29 3580.01 3579.94 96.7 303550.96 3550.89 99.9 31 3470.87 3471.16 99.9 32 3293.67 3293.80 99.0 333481.90 3481.80 95.8 34 3578.90 3578.70 98.6 35 3564.95 3564.30 99.9 363512.91 3512.54 99.9 37 3498.89 3498.95 99.9 38 3484.90 3484.75 99.9 393498.93 3498.87 96.8 40 3324.68 3324.38 98.6

A compound of the present invention can be tested for activity as aGLP-1 binding compound according to the following procedure.

Cell Culture:

RIN 5F rat insulinoma cells (ATCC-# CRL-2058, American Type CultureCollection, Manassas, Va.), expressing the GLP-1 receptor, were culturedin Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal calfserum, and maintained at about 37° C. in a humidifed atmosphere of 5%CO₂/95% air.

Radioligand Binding:

Membranes were prepared for radioligand binding studies byhomogenization of the RIN cells in 20 ml of ice-cold 50 mM Tris-HCl witha Brinkman Polytron (Westbury, N.Y.) (setting 6, 15 sec). Thehomogenates were washed twice by centrifugation (39,000 g/10 min), andthe final pellets were resuspended in 50 mM Tris-HCl, containing 2.5 mMMgCl₂, 0.1 mg/ml bacitracin (Sigma Chemical, St. Louis, Mo.), and 0.1%BSA. For assay, aliquots (0.4 ml) were incubated with 0.05 nM(¹²⁵I)GLP-1(7-36) (˜2200 Ci/mmol, New England Nuclear, Boston, Mass.),with and without 0.05 ml of unlabeled competing test peptides. After a100 min incubation (25° C.), the bound (¹²⁵I)GLP-1(7-36) was separatedfrom the free by rapid filtration through GF/C filters (Brandel,Gaithersburg, Md.), which had been previously soaked in 0.5%polyethyleneimine. The filters were then washed three times with 5 mlaliquots of ice-cold 50 mM Tris-HCl, and the bound radioactivity trappedon the filters was counted by gamma spectrometry (Wallac LKB,Gaithersburg, Md.). Specific binding was defined as the total(¹²⁵I)GLP-1(7-36) bound minus that bound in the presence of 1000 nMGLP1(7-36) (Bachem, Torrence, Calif.).

1. A compound according to the formula: (Aib^(8,35), Arg^(26,34), Phe³¹,Pro³⁷, Ser^(38,39))hGLP-1(7-39)-NH₂; (SEQ ID NO:1) (Aib^(8,35,37),Arg^(26,34), Phe³¹, Asn³⁸)hGLP-1(7-38)-NH₂; (SEQ ID NO:2)(Aib^(8,35,37), Arg^(26,34), Phe³¹, Ser³⁸)hGLP-1(7-38) NH₂; (SEQ IDNO:3) (Aib^(8,35,37), Gaba³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:4)(Aib^(8,35,37), Arg^(26,34), Phe³¹, His³⁸)hGLP-1(7-38) NH₂; (SEQ IDNO:5) (Aib^(8,35), Arg^(26,34), Phe³¹, β-Ala³⁷, His³⁸)hGLP-1(7-38) NH₂;(SEQ ID NO:6) (Aib^(8,35,37), Arg^(26,34), D-His³⁸)hGLP-1(7-38) NH₂;(SEQ ID NO:7) (Aib^(8,35,37), β-Ala³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:8)(Aib^(8,35), Arg^(26,34), β-Ala³⁷, His³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:9)(Aib^(8,35,37), Arg^(26,34), Phe³¹, Gly³⁸)hGLP-1(7-38) NH₂; (SEQ IDNO:10) (Aib^(8,35,37), Arg^(26,34), Gly³⁸)hGLP-1(7-38) NH₂; (SEQ IDNO:11) (Aib^(8,35,37), Arg^(26,34), β-Ala³⁸)hGLP-1(7-38) NH₂; (SEQ IDNO:12) (Aib^(8,35,37), Arg^(26,34), Gaba³⁸)hGLP-1(7-38) NH₂; (SEQ IDNO:13) (Aib^(8,35,37), Arg³⁴, Phe³¹, His³⁸)hGLP-1(7-38) NH₂; (SEQ IDNO:14) (Aib^(8,35,37), Arg^(26,34), His³⁸)hGLP-1(7-38) NH₂; (SEQ IDNO:15) (Aib^(8,35,37), Arg^(26,34), Phe³¹, Gaba³⁸)hGLP-1(7-38) NH₂; (SEQID NO:16) (Aib^(8,35,37), Arg³⁴, Phe³¹, D-His³⁸)hGLP-1(7-38) NH₂; (SEQID NO:19) (Aib^(8,35,37), Arg³⁴, Phe³¹, Gly³⁸)hGLP-1(7-38) NH₂; (SEQ IDNO:20) (Aib^(8,35,37), Gly³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:21)(Aib^(8,35,37), Arg^(26,34), Phe³¹, D-His³⁸)hGLP-1(7-38) NH₂; (SEQ IDNO:22) (Aib^(8,35), Arg^(26,34), Phe³¹, β-Ala³⁷, D-His³⁸)hGLP-1(7-38)NH₂; (SEQ ID NO:23) (Aib^(8,35,37), Arg^(26,34), Phe³¹,β-Ala³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:24) (Aib^(8,35), Arg^(26,34),Phe³¹, β-Ala^(37,38))hGLP-1(7-38) NH₂; (SEQ ID NO:25) (Aib^(8,35,37),Arg³⁴, Phe³¹, β-Ala³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:26) or(Aib^(8,35,37), Arg³⁴, Phe³¹, Gaba³⁸)hGLP-1(7-38) NH₂; (SEQ ID NO:27) ora pharmaceutically acceptable salt thereof.
 2. A pharmaceuticalcomposition comprising a compound according to claim 1 or apharmaceutically acceptable salt thereof and a pharmaceuticallyacceptable carrier or diluent.